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CHEM 222
Karine Auclair

Chemistry 212 Fall 2013 Experiment 6: Column Chromatography and TLC of Spinach – Evaluation Summary Last Name:Atique First Name:Anika Date: 12/3/2013 Lab Day/TA/Group: Wednesday, Patrick Julien, Group C Lab Partner(s): Labs reports must be typed and chemical structures must be drawn with ChemDraw. Report must not exceed five pages (including this page). Page limit does not include any attached spectra. ** Deductions for hand written report/structures and exceeding page limit ** Report Breakdown Intro/Results: _________________ /10 Discussion: _________________ /10 Report Total: _________________ /20 Other Lab Marks Performance: _________________ /6 Prelab Quiz: _________________ /3 TAComments on Report/Performance Report assessment TA(name): TA(signature): Performance assessment TA(name): TA(signature): 1 of 6 Chemistry 212 Fall 2013 Experiment 6: Column Chromatography and TLC of Spinach – Lab Report Guidelines Include the following in your report (in the following order, use provided table): 1. Title, Date, Your Name, Your Day/Time/TA AtiqueAnika. Lab day: Wednesday, TA: Patrick Julien, Group: C 2. Objective: (1 mark) What is the purpose of this experiment? The purpose of this experiment is to extract and separate the different pigments of the spinach extract. 3. Introduction: (2 marks) Brief description of the concept/reaction studied – extraction, column chromatography – and why it is important The extraction process is efficient but is indiscriminate so we end up with a mixture of pigments. This is why separation by column chromatography is important to be able to identify the different pigments present. Extraction is done by grinding the spinach leaves to obtain a pigment mixture and then using a separating funnel to isolate only the pigment extract from the water present. Column chromatography is used to separate the pigments based on their polarity. The more polar pigments tend to associate more with the polar silica stationary phase of the column while the less polar components associate more with the non polar solvent. This difference in the degrees of association separates the pigments into clear layers. 4. Procedure: (2 marks) Summary of lab procedure – what YOU did, include KEY observations – be concise! Preparing Column:Added a cotton blob and some sand to the column. Dissolved silica in eluent and added this to the column in turns while draining the eluent to make the stationary phase.Add a final top layer of sand to keep the silica in place while always ensuring there was enough eluent present to keep the silica wet. Stopped the stopcock once the eluent level reached the top sand layer. Development and elution of column: Added the pigment extract to the column using a pipette and used eluent until the green layer of pigment dropped below the sand. The addition of eluent was continued to rinse the column until it was colorless. The eluent was added in turns and drained off until all the colored band appeared. Different bands were observed- cloudy white just below the level of the top sand layer, then in descending manner, bluish green, dark green, blue green, cloudy white and finally yellow. The 3 different pigment were then drained into individual beakers (blue-green, yellow and green). TLC: 3 different TLC plates were set up using different ratios of acetone and ether for the mobile phase. 9:1, 8:2 and 7:3. The three different pigments along with the crude sample was spotted onto the plate and the components were allowed to separate. In the 9:1 plate The crude separated into one spot and only the yellow pigment gave a spot. In the 8:2 plate the crude gave three spots 2 Chemistry 212 Fall 2013 and the 3 pigments each gave an individual spot. In the 7:3 plate The crude gave 3 spots and the individual pigments each gave their ow
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